EP0014490B1 - Process for the separation of poly-beta-hydroxybutyrates from a biomass - Google Patents

Process for the separation of poly-beta-hydroxybutyrates from a biomass Download PDF

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Publication number
EP0014490B1
EP0014490B1 EP80200032A EP80200032A EP0014490B1 EP 0014490 B1 EP0014490 B1 EP 0014490B1 EP 80200032 A EP80200032 A EP 80200032A EP 80200032 A EP80200032 A EP 80200032A EP 0014490 B1 EP0014490 B1 EP 0014490B1
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Prior art keywords
extraction
biomass
hydroxybutyrates
chloropropanes
chloroethanes
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German (de)
French (fr)
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EP0014490A1 (en
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Noel Vanlautem
Jacques Gilain
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Solvay SA
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Solvay SA
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/62—Carboxylic acid esters
    • C12P7/625—Polyesters of hydroxy carboxylic acids

Definitions

  • the present invention relates to a process for the separation of poiy-p-hydroxybutyrates from a biomass by extraction using liquid halogenated solvents.
  • microorganisms are capable of synthesizing poiy-p-hydroxybutyrates whose main function in these microorganisms seems to be the storage of energy in the form of carbonaceous matter.
  • These poiy-p-hydroxybutyrates constitute an interesting raw material on the Industrial level.
  • Various techniques have already been considered for separating them from biomass.
  • it has already been proposed to extract them from the biomass by means of low-boiling liquid halogenated solvents such as chloroform and methylene chloride.
  • the extraction is generally carried out at the normal boiling point of the solvent or at a lower temperature.
  • the, mining requires very long processing times and is accompanied by a depolymerization of poly- ⁇ -hydroxybutyrate although it operates at low temperature.
  • solvents such as cyclic carbonate of 1,2-ethanediol (ethylene carbonate) or 1,2-propanediol (propylene carbonate) at higher temperatures.
  • the present invention aims to provide a method which no longer has any of the drawbacks of the known methods.
  • the present invention relates to a process for the separation of poly-p-hydroxybutyrates from a biomass by extraction using liquid halogenated solvents, according to which liquid halogenated solvents chosen from chloroethanes and chloropropanes are used.
  • chloroethanes and chloropropanes is intended to denote all the compounds derived from ethane or propane in which one or more hydrogen atoms has or have been substituted by the chlorine atom, as well as the chloroethanes and chloropropanes of which certain hydrogen atoms have also been substituted by halogen atoms such as iodine and bromine and, preferably, fluorine. It is however preferred to use chloroethanes and chloropropanes which are unsubstituted and which contain in the molecule only chlorine, carbon and hydrogen atoms.
  • the liquid medium used for the extraction according to the invention although consisting essentially of chloroethanes and chloropropanes, may contain small amounts of other liquids. However, it is preferred that it consists of chloroethanes and chloropropanes in an amount of 90% by weight, and more particularly 99% by weight, at least.
  • solvents used according to the invention can be used alone or in combination.
  • the simplest execution consists in the use of a single solvent possibly in the impure form commonly called "of technical quality".
  • solvents are chosen which have a boiling point, measured at atmospheric pressure, of between 65 ° C. and 170 ° C.
  • the most particularly preferred solvents are unsubstituted chloroethanes and chloropropanes, the carbon atoms of which contain at least one chlorine atom are also substituted with at least one hydrogen atom.
  • the solvents which give the best results are 1,2-dichloroethane, 1,1,2-trichloroethane, 1,1,2,2-tetrachloroethane and 1,2,3-trichloropropane. The best results have been obtained with 1,2-dichloroethane and 1,1,2-trichloroethane.
  • the biomasses which can be treated according to the invention can be obtained from microorganisms of various origins and in particular from bacteria as it has been described in Angew. Chemie 1962, 74 1967g, Nr. 10 p. 342-346.
  • the selection of microorganisms is generally made on the basis of the relative amount of poly-p-hydroxybutyrates contained in the microorganism as well as according to the growth rate of the microorganism and its speed of synthesis of poly- ⁇ -hydroxybutyrates.
  • Microorganisms can be treated directly with the extraction solvent in their culture medium.
  • a preferred embodiment consists of microorganisms which have been, prior to extraction, separated from their culture medium. This separation can be carried out by any means known for this purpose.
  • An elegant way of proceeding consists in centrifuging the culture medium so as to be able to easily separate the microorganisms from the medium and subsequently lyophilizing the isolated microorganisms so as to obtain a biomass which can be easily treated with the extraction solvent.
  • the relationship between the amount of biomass used and the extraction solvent used is not critical in itself. We generally work with weight ratios between 1 to 1 and 1 to 100. Usually, we use ratios between 1 to 2 and 1 to 50 and finally we operate preferably with ratios between 1 to 5 and 1 to 10. The choice of this ratio is influenced by various parameters such as the nature of the biomass to be treated, the temperature, the number of extracts and the yield desired by extraction. In addition, the elimination of cellular residues is all the easier when there is less biomass in the extraction medium.
  • the temperature at which the extraction is carried out is chosen according to the nature of the chloroethanes and chloropropanes used. Generally suitable temperatures are between 50 ° C and the boiling point, measured at atmospheric pressure, of the solvent. It has been found that the optimum operating temperatures are usually below 130 ° C. They are from preferably between 60 and 90 ° C. In the case of 1,2-dichloroethane, good results have been obtained between 60 and 80 ° C.
  • the pressure at which the extraction is carried out is not critical and is generally between 0.1 and 10 kg / cm 2 [98-9807 mbar].
  • the extraction operation can be carried out in any apparatus designed for this purpose.
  • the operation can be carried out continuously or discontinuously with circulations of the biomass and extraction solvents circulating in the same direction or in opposite directions.
  • the solvent can be freed of compounds that are not soluble in the medium, such as the cell membranes, of the extracted biomass.
  • any known means can be used. Usually this operation is carried out by one or more filtrations.
  • the extracted poly-p-hydroxybutyrates can be separated from the extraction medium by any method known for this purpose. Thus, they can be separated by evaporation of the solvent or even by simple addition of a precipitating agent.
  • precipitating agents which can be used, mention may be made of the non-solvent compounds of poly-p-hydroxybutyrates such as petroleum ether, unsubstituted aliphatic hydrocarbons, aromatic solvents including benzene or aliphatic alcohols.
  • Preferred precipitating agents are aliphatic alcohols and more particularly, for economic reasons, methanol and ethanol.
  • the separated poly-p-hydroxybutyrates according to the invention can then be purified by one or more washings with non-solvents such as those mentioned above and be dried, after which they are in the form of a white polymeric mass. Recovery and purification operations are usually carried out at room temperature.
  • Poy-p-hydroxybutyrates are polymers having many applications and in particular in surgery where they can be used in the form of wires because they are easily sterilizable.
  • these polymers can be shaped by the various known molding techniques for making prostheses. We can still spin or extrude them according to the usual methods:
  • the extraction yield is given by the weight of the product extracted over the weight of biomass used and the molecular weight by weight of polymer is calculated from the measurement of the intrinsic viscosity of a chloroform solution according to the formula recommended in the article published in Makromol. Chem. Flight. 176, 1975 p. 2662. Values obtained are collated in Table 1 below.
  • Example 1 is repeated, but substituting tetrachloromethane (example 2) and trichlorethylene (example 3) respectively for chloroform. The results obtained are collated in Table 1.
  • Example 4 1,2-dichloroethane
  • Example 5 1,1,2-Trichloroethane
  • Example 6 1,1,2,2-tetrachloroethane
  • Example 1 is repeated with chloroform as extraction solvent but while carrying out the extraction for a period of 3 hours.
  • the results obtained are shown in Table 1 below.
  • Example 2 The procedure is as in Example 1 but instead of working at 60 ° C, the extraction is carried out at 120 ° C and with propylene carbonate instead of chloroform. The observed yield and the molecular weight of the extracted polymer are listed in Table 2 below.
  • Example 8 The procedure is as in Example 8 but replacing the propylene carbonate with 1.1,2,2-tetrachloroethane (Example 9) and with 1,2,3-Trichloropropane (Example 10). The results are collated in Table 2.
  • the invention makes it possible to obtain, at the same temperature, extraction yields which are of the same order of magnitude as those obtained with propylene carbonate while not causing such significant depolymerization of the extracted polymer.

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  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Extraction Or Liquid Replacement (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Polyesters Or Polycarbonates (AREA)
  • Removal Of Specific Substances (AREA)
  • Epoxy Compounds (AREA)

Abstract

A process for separating poly- beta -hydroxybutyrates from a biomass by extraction using at least one liquid halogenated solvent selected from the group consisting of chloroethanes and chloropropanes.

Description

La présente invention concerne un procédé de séparation de poiy-p-hydroxybutyrates d'une biomasse par extraction au moyen de solvants halogénés liquides.The present invention relates to a process for the separation of poiy-p-hydroxybutyrates from a biomass by extraction using liquid halogenated solvents.

De nombreux microorganismes sont capables de synthétiser des poiy-p-hydroxybutyrates dont la fonction principale dans ces microorganismes semble être le stockage de l'énergie sous forme de matière carbonée. Ces poiy-p-hydroxybutyrates constituent une matière première intéressante sur le plan Industriel. Diverses techniques ont déjà été envisagées pour les séparer des biomasses. Ainsi, on a déjà proposé de les extraire de la biomasse au moyen de solvants halogénés liquides à bas point d'ébullition tels que le chloroforme et le chlorure de méthylène. Dans ces procédés connus, l'extraction est généralement réalisée à la température normale d'ébullition du solvant ou à une température inférieure. Ainsi réalisée, l,extraction nécessite des durées de traitement très longues et est accompagnée d,une dépolymérisation des poly-β-hydroxybutyrates bien que l,on opère à basse température.Many microorganisms are capable of synthesizing poiy-p-hydroxybutyrates whose main function in these microorganisms seems to be the storage of energy in the form of carbonaceous matter. These poiy-p-hydroxybutyrates constitute an interesting raw material on the Industrial level. Various techniques have already been considered for separating them from biomass. Thus, it has already been proposed to extract them from the biomass by means of low-boiling liquid halogenated solvents such as chloroform and methylene chloride. In these known processes, the extraction is generally carried out at the normal boiling point of the solvent or at a lower temperature. Thus conducted, the, mining requires very long processing times and is accompanied by a depolymerization of poly-β-hydroxybutyrate although it operates at low temperature.

Dans le but de réduire les durées d'extraction et d'augmenter les rendements, on a proposé de mettre en oeuvre des solvants tels que le carbonate cyclique de 1,2-éthanediol (carbonate d'éthylène) ou de 1,2-propanediol (carbonate de propylène) à des températures plus élevées. Ces procédés permettent effectivement d,atteindre les buts visés mais ils engendrent une dépolymérisation importante des poly-β-hydroxybutyrates.In order to reduce extraction times and increase yields, it has been proposed to use solvents such as cyclic carbonate of 1,2-ethanediol (ethylene carbonate) or 1,2-propanediol (propylene carbonate) at higher temperatures. These methods allow to effectively, meet those goals but they generate a significant depolymerization of poly-β-hydroxybutyrate.

La présente invention vise à fournir un procédé qui ne présente plus aucun des inconvénients des procédés connus.The present invention aims to provide a method which no longer has any of the drawbacks of the known methods.

La présente invention concerne à cet effet un procédé de séparation de poly-p-hydroxybutyrates d'une biomasse par extraction au moyen de solvants halogénés liquides, selon lequel on utilise des solvants halogénés liquides choisis parmi les chloroéthanes et les chloropropanes.To this end, the present invention relates to a process for the separation of poly-p-hydroxybutyrates from a biomass by extraction using liquid halogenated solvents, according to which liquid halogenated solvents chosen from chloroethanes and chloropropanes are used.

Par chloroéthanes et chloropropanes, on entend désigner tous les composés dérivés de l'éthane ou du propane dans lesquels un ou plusieurs atomes d'hydrogène a ou ont été substitué(s) par l'atome de chlore ainsi que les chloroéthanes et les chloropropanes dont certains atomes d'hydrogène ont en outre été substitués par des atomes d'halogène tels que l'iode et le brome et, de préférence, le fluor. On préfère toutefois utiliser des chloroéthanes et des chloropropanes qui ne sont pas substitués et qui comportent dans la molécule uniquement des atomes de chlore, de carbone et d'hydrogène.The term “chloroethanes and chloropropanes” is intended to denote all the compounds derived from ethane or propane in which one or more hydrogen atoms has or have been substituted by the chlorine atom, as well as the chloroethanes and chloropropanes of which certain hydrogen atoms have also been substituted by halogen atoms such as iodine and bromine and, preferably, fluorine. It is however preferred to use chloroethanes and chloropropanes which are unsubstituted and which contain in the molecule only chlorine, carbon and hydrogen atoms.

Le milieu liquide utilisé pour l'extraction selon l'invention, bien que constitué essentiellement de chloroéthanes et de chloropropanes, peut contenir de faibles quantités d'autres liquides. On préfère cependant qu'il soit constitué de chloroéthanes et de chloropropanes à raison de 90 % en poids, et plus particulièrement 99 % en poids, au moins.The liquid medium used for the extraction according to the invention, although consisting essentially of chloroethanes and chloropropanes, may contain small amounts of other liquids. However, it is preferred that it consists of chloroethanes and chloropropanes in an amount of 90% by weight, and more particularly 99% by weight, at least.

Les solvants utilisés selon l'invention peuvent être utilisés seuls ou en combinaison. L'exécution la plus simple consiste dans l'emploi d'un seul solvant éventuellement sous la forme impure appelée couramment « de qualité technique •. De préférence, on choisit des solvants qui possèdent un point d'ébullition, mesuré à pression atmosphérique, compris entre 65 °C et 170 °C.The solvents used according to the invention can be used alone or in combination. The simplest execution consists in the use of a single solvent possibly in the impure form commonly called "of technical quality". Preferably, solvents are chosen which have a boiling point, measured at atmospheric pressure, of between 65 ° C. and 170 ° C.

Les solvants tout particulièrement préférés sont les chloroéthanes et chloropropanes non substitués dont les atomes carbone porteurs d'au moins un atome de chlore sont également substitués par au moins un atome d'hydrogène. Dans cette catégorie les solvants qui donnent les meilleurs résultats sont le 1,2-dichloroéthane, le 1,1,2-trichloroéthane, le 1,1,2,2-tétrachloroéthane et le 1,2,3-trichloropropane. Les meilleurs résultats ont été obtenus avec le 1,2-dichloroéthane et le 1,1,2-trichloroéthane.The most particularly preferred solvents are unsubstituted chloroethanes and chloropropanes, the carbon atoms of which contain at least one chlorine atom are also substituted with at least one hydrogen atom. In this category the solvents which give the best results are 1,2-dichloroethane, 1,1,2-trichloroethane, 1,1,2,2-tetrachloroethane and 1,2,3-trichloropropane. The best results have been obtained with 1,2-dichloroethane and 1,1,2-trichloroethane.

Les biomasses que l'on peut traiter selon l'invention peuvent être obtenues à partir de microorganismes d'origines diverses et notamment à partir de bactéries comme il a été décrit dans Angew. Chemie 1962, 74 Jahrg, Nr. 10 p. 342-346. La sélection des microorganismes se fait en général sur la base de la quantité relative de poly-p-hydroxybutyrates contenue dans le microorganisme ainsi qu'en fonction de la vitesse de croissance du microorganisme et de sa vitesse de synthèse des poly-β-hydroxybutyrates.The biomasses which can be treated according to the invention can be obtained from microorganisms of various origins and in particular from bacteria as it has been described in Angew. Chemie 1962, 74 Jahrg, Nr. 10 p. 342-346. The selection of microorganisms is generally made on the basis of the relative amount of poly-p-hydroxybutyrates contained in the microorganism as well as according to the growth rate of the microorganism and its speed of synthesis of poly-β-hydroxybutyrates.

Les microorganismes, qu'ils soient cultivés artificiellement ou non, peuvent être traités directement par le solvant d'extraction dans leur milieu de culture. Une exécution préférée consiste toutefois à partir de microorganismes qui ont été, préalablement à l'extraction, séparés de leur milieu de culture. Cette séparation peut être effectuée par tous les moyens connus à cet effet. Une façon élégante de procéder consiste à centrifuger le milieu de culture de façon à pouvoir aisément séparer les microorganismes du milieu et à lyophiliser subséquement les microorganismes isolés de façon à obtenir une biomasse qui peut être traitée aisément par le solvant d'extraction.Microorganisms, whether artificially cultivated or not, can be treated directly with the extraction solvent in their culture medium. A preferred embodiment, however, consists of microorganisms which have been, prior to extraction, separated from their culture medium. This separation can be carried out by any means known for this purpose. An elegant way of proceeding consists in centrifuging the culture medium so as to be able to easily separate the microorganisms from the medium and subsequently lyophilizing the isolated microorganisms so as to obtain a biomass which can be easily treated with the extraction solvent.

Le rapport entre la quantité de la biomasse mise en oeuvre et le solvant d'extraction utilisé n'est pas critique en soi. On travaille en général avec des rapports en poids compris entre 1 à 1 et 1 à 100. Habituellement, on emploie des rapports compris entre 1 à 2 et 1 à 50 et enfin on opère de préférence avec des rapports compris entre 1 à 5 et 1 à 10. Le choix de ce rapport est influencé par divers paramètres tels que la nature de la biomasse à traiter, la température, le nombre d'extractions et le rendement souhaité par extraction. En outre, l'élimination des résidus cellulaires est d'autant plus facile qu'il y a moins de biomasse dans le milieu d'extraction.The relationship between the amount of biomass used and the extraction solvent used is not critical in itself. We generally work with weight ratios between 1 to 1 and 1 to 100. Usually, we use ratios between 1 to 2 and 1 to 50 and finally we operate preferably with ratios between 1 to 5 and 1 to 10. The choice of this ratio is influenced by various parameters such as the nature of the biomass to be treated, the temperature, the number of extracts and the yield desired by extraction. In addition, the elimination of cellular residues is all the easier when there is less biomass in the extraction medium.

La température à laquelle est effectuée l'extraction est choisie en fonction de la nature des chloroéthanes et chloropropanes utilisés. Les températures qui conviennent bien en général sont situées entre 50 °C et le point d'ébullition, mesuré sous pression atmosphérique, du solvant. On a constaté que les températures optimales d'opération se situent habituellement en dessous de 130 °C. Elles sont de préférence comprises entre 60 et 90 °C. Dans le cas du 1,2-dichloroéthane, de bons résultats ont été obtenus entre 60 et 80 °C.The temperature at which the extraction is carried out is chosen according to the nature of the chloroethanes and chloropropanes used. Generally suitable temperatures are between 50 ° C and the boiling point, measured at atmospheric pressure, of the solvent. It has been found that the optimum operating temperatures are usually below 130 ° C. They are from preferably between 60 and 90 ° C. In the case of 1,2-dichloroethane, good results have been obtained between 60 and 80 ° C.

La pression à laquelle est effectuée l'extraction n'est pas critique et est généralement comprise entre 0,1 et 10 kg/cm2 [98-9807 mbar].The pressure at which the extraction is carried out is not critical and is generally between 0.1 and 10 kg / cm 2 [98-9807 mbar].

L'opération d'extraction peut être réalisée dans tout appareillage conçu à cet effet. On peut effectuer l'opération en continu ou en discontinu avec des circulations des biomasses et des solvants d'extraction circulant dans le même sens ou dans des sens opposés.The extraction operation can be carried out in any apparatus designed for this purpose. The operation can be carried out continuously or discontinuously with circulations of the biomass and extraction solvents circulating in the same direction or in opposite directions.

Après l'opération d'extraction le solvant peut être débarrassé des composés non solubles dans le milieu tels que les membranes cellulaires de la biomasse extraite. Pour ce faire, on peut utiliser n'importe quel moyen connu. Habituellement cette opération est effectuée par une ou plusieurs filtrations.After the extraction operation, the solvent can be freed of compounds that are not soluble in the medium, such as the cell membranes, of the extracted biomass. To do this, any known means can be used. Usually this operation is carried out by one or more filtrations.

Les poly-p-hydroxybutyrates extraits peuvent être séparés du milieu d'extraction par n'importe quelle méthode connue à cet effet. Ainsi, on peut les séparer par évaporation du solvant ou encore par simple addition d'un agent précipitant. Comme agents précipitants utilisables, on peut citer les composés non solvants des poly-p-hydroxybutyrates tels que l'éther de pétrole, les hydrocarbures aliphatiques non substitués, les solvants aromatiques dont le benzène ou encore les alcools aliphatiques. Des agents précipitants préférés sont les alcools aliphatiques et plus particulièrement, pour des raisons économiques, le méthanol et l'éthanol.The extracted poly-p-hydroxybutyrates can be separated from the extraction medium by any method known for this purpose. Thus, they can be separated by evaporation of the solvent or even by simple addition of a precipitating agent. As precipitating agents which can be used, mention may be made of the non-solvent compounds of poly-p-hydroxybutyrates such as petroleum ether, unsubstituted aliphatic hydrocarbons, aromatic solvents including benzene or aliphatic alcohols. Preferred precipitating agents are aliphatic alcohols and more particularly, for economic reasons, methanol and ethanol.

Les poly-p-hydroxybutyrates séparés selon l'invention peuvent être ensuite purifiés par un ou plusieurs lavages avec des non solvants tels que ceux cités ci-avant et être séchés, après quoi ils se présentent sous la forme d'une masse polymérique blanche. Les opérations de récupération et de purification se font habituellement à température ambiante.The separated poly-p-hydroxybutyrates according to the invention can then be purified by one or more washings with non-solvents such as those mentioned above and be dried, after which they are in the form of a white polymeric mass. Recovery and purification operations are usually carried out at room temperature.

Les poiy-p-hydroxybutyrates sont des polymères ayant de nombreuses applications et notamment en chirurgie où ils peuvent être utilisés sous forme de fils car ils sont aisément stérilisables. En outre, ces polymères peuvent être mis en forme par les différentes techniques connues de moulage pour faire des prothèses. On peut encore les filer ou les extruder suivant les méthodes habituelles:Poy-p-hydroxybutyrates are polymers having many applications and in particular in surgery where they can be used in the form of wires because they are easily sterilizable. In addition, these polymers can be shaped by the various known molding techniques for making prostheses. We can still spin or extrude them according to the usual methods:

Les exemples suivants servent à illustrer l'invention.The following examples serve to illustrate the invention.

Exemple 1 (de comparaison)Example 1 (comparison)

Dans un ballon de 250 ml muni d'un agitateur et d'un réfrigérant à eau on introduit 100 ml de chloroforme que l'on porte à 60 °C. On introduit ensuite 16,0 g d'une biomasse séchée lyophilisée constituée de bactéries de la souche ALCALIGENES EUTROPHUS, délitée au mortier avant l'emploi, et dont la teneur en eau superficielle est de 23,2 g/kg. Après 1 heure, on prélève la solution et on filtre sous pression à chaud sur une toile en tissu afin d'éliminer les restes solides de la biomasse. Le polymère est précipité par addition d'éthanol en excès, à température ambiante, à la solution obtenue. Le précipité est filtré sous vide sur un filtre avec verre fritté et lavé avec de l'éthanol. Le polymère est ensuite séché jusqu'à poids constant sous vide. Le rendement de l'extraction est donné par le poids du produit extrait sur le poids de biomasse mis en oeuvre et le poids moléculaire en poids de polymère est calculé à partir de la mesure de la viscosité intrinsèque d'une solution chloroformique selon la formule

Figure imgb0001
préconisée dans l'article paru dans Makromol. Chem. Vol. 176, 1975 p. 2662. Des valeurs obtenues sont rassemblées au Tableau 1 ci-après.100 ml of chloroform are introduced into a 250 ml flask fitted with an agitator and a water cooler, which is brought to 60 ° C. Then introduced 16.0 g of a lyophilized dried biomass consisting of bacteria of the strain ALCALIGENES EUTROPHUS, disintegrated in mortar before use, and whose surface water content is 23.2 g / kg. After 1 hour, the solution is taken and filtered under hot pressure on a cloth to remove solid residues from the biomass. The polymer is precipitated by adding excess ethanol, at room temperature, to the solution obtained. The precipitate is filtered under vacuum on a filter with sintered glass and washed with ethanol. The polymer is then dried to constant weight under vacuum. The extraction yield is given by the weight of the product extracted over the weight of biomass used and the molecular weight by weight of polymer is calculated from the measurement of the intrinsic viscosity of a chloroform solution according to the formula
Figure imgb0001
 recommended in the article published in Makromol. Chem. Flight. 176, 1975 p. 2662. Values obtained are collated in Table 1 below.

Exemples 2 et 3 (de comparaison)Examples 2 and 3 (for comparison)

On répète l'exemple 1 mais en substituant respectivement du tétrachlorométhane (exemple 2) et du trichloroéthylène (exemple 3) au chloroforme. Les résultats obtenus sont rassemblés au Tableau 1.Example 1 is repeated, but substituting tetrachloromethane (example 2) and trichlorethylene (example 3) respectively for chloroform. The results obtained are collated in Table 1.

Exemples 4, 5 et 6Examples 4, 5 and 6

On opère de nouveau dans des conditions identiques à celles de l'exemple 1 mais on effectue l'opération d'extraction avec du 1,2-dichloroéthane (exemple 4), du 1,1,2-trichloroéthane (exemple 5) et du 1,1,2,2-tétrachloroéthane (exemple 6). Les résultats obtenus sont également repris au Tableau 1 ci-après.The operation is again carried out under conditions identical to those of Example 1, but the extraction operation is carried out with 1,2-dichloroethane (Example 4), 1,1,2-Trichloroethane (Example 5) and 1,1,2,2-tetrachloroethane (Example 6). The results obtained are also given in Table 1 below.

Exemple 7 (de comparaison)Example 7 (for comparison)

L'exemple 1 est répété avec du chloroforme comme solvant d'extraction mais en opérant l'extraction pendant une durée de 3 heures. Les résultats obtenus sont repris au Tableau 1 ci-dessous.

Figure imgb0002
Example 1 is repeated with chloroform as extraction solvent but while carrying out the extraction for a period of 3 hours. The results obtained are shown in Table 1 below.
Figure imgb0002

On peut donc conclure des résultats rassemblés au Tableau 1 que les rendements observés par extraction selon l'invention sont nettement plus élevés qu'avec d'autres solvants halogénés dans les mêmes conditions opératoires. Ces rendements sont même plus élevés que ceux obtenus après une durée d'extraction triple avec un solvant halogéné typique de l'art antérieur.It can therefore be concluded from the results collected in Table 1 that the yields observed by extraction according to the invention are significantly higher than with other halogenated solvents under the same operating conditions. These yields are even higher than those obtained after a triple extraction time with a halogenated solvent typical of the prior art.

En outre on peut voir que dans les exemples 4, 5, 6 les poids moléculaires des polymères séparés selon l'invention sont nettement supérieurs à ceux obtenus à la même température avec le chloroforme.Furthermore, it can be seen that in Examples 4, 5, 6 the molecular weights of the polymers separated according to the invention are much higher than those obtained at the same temperature with chloroform.

Exemple 8 (de comparaison)Example 8 (for comparison)

On opère comme dans l'exemple 1 mais au lieu de travailler à 60 °C on effectue l'extraction à 120 °C et avec du carbonate de propylène au lieu de chloroforme. Le rendement observé et le poids moléculaire du polymère extrait sont repris au Tableau 2 ci-après.The procedure is as in Example 1 but instead of working at 60 ° C, the extraction is carried out at 120 ° C and with propylene carbonate instead of chloroform. The observed yield and the molecular weight of the extracted polymer are listed in Table 2 below.

Exemples 9 et 10Examples 9 and 10

On travaille comme dans l'exemple 8 mais en remplaçant le carbonate de propylène par du 1.1,2,2-tétrachloroéthane (exemple 9) et par du 1,2,3-trichloropropane (exemple 10). Les résultats sont rassemblés au Tableau 2.

Figure imgb0003
The procedure is as in Example 8 but replacing the propylene carbonate with 1.1,2,2-tetrachloroethane (Example 9) and with 1,2,3-Trichloropropane (Example 10). The results are collated in Table 2.
Figure imgb0003

On peut donc déduire de ces résultats que l'invention permet d'obtenir, à la même température, des rendements d'extraction qui sont du même ordre de grandeur que ceux obtenus avec le carbonate de propylène tout en ne provoquant pas une dépolymérisation aussi importante du polymère extrait.It can therefore be deduced from these results that the invention makes it possible to obtain, at the same temperature, extraction yields which are of the same order of magnitude as those obtained with propylene carbonate while not causing such significant depolymerization of the extracted polymer.

En outre, on peut déduire, de la comparaison des résultats des Tableaux 1, et 2, que l'invention permet d'obtenir des rendements d'extraction meilleurs ou tout au moins égaux à des températures de 60 °C comparés aux rendements des procédés connus à 120°C sans provoquer une dépolymérisation aussi importante.In addition, it can be deduced from the comparison of the results of Tables 1 and 2 that the invention makes it possible to obtain better or at least equal extraction yields at temperatures of 60 ° C. compared to the process yields. known at 120 ° C without causing such significant depolymerization.

Claims (5)

1. Process for separating poly-p-hydrox-ybutyrates from a biomass by extraction using liquid halogenated solvents, characterised in that the liquid halogenated solvents are chosen from amongst chloroethanes and chloropropanes.
2. Process according to Claim 1, characterised in that the liquid halogenated solvents are chosen from amongst chloroethanes and chloropropanes having a boiling point of between 65 °C and 170 °C.
3. Process accordmg to Claim 1 or 2, characterised in that the liquid halogenated solvents are chosen from amongst chloroethanes and chloropropanes which possess carbon atoms carrying at least one chlorine atom and also at least one hydrogen atom.
4. Process according to any one of Claims 1 to 3, characterised in that the liquid halogenated solvents are chosen from amongst 1,2-dichloroethane, 1,1,2-trichloroethane, 1,1,2,2-tetrachloroethane and 1,2,3-trichloropropane.
5. Process according to any one of Claims 1 to 4, characterised in that the extraction is carried out at between 60 °C and 90 °C.
EP80200032A 1979-01-22 1980-01-14 Process for the separation of poly-beta-hydroxybutyrates from a biomass Expired EP0014490B1 (en)

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AT80200032T ATE1649T1 (en) 1979-01-22 1980-01-14 PROCESS FOR THE SEPARATION OF POLY-BETAHYDROXYBUTYRATES FROM A BIOMASS.

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FR7901862 1979-01-22
FR7901862A FR2446859A1 (en) 1979-01-22 1979-01-22 PROCESS FOR SEPARATING POLY-B-HYDROXYBUTYRATES FROM A BIOMASS

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JPS5599195A (en) 1980-07-28
FR2446859A1 (en) 1980-08-14
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